The mobility of troponin C and troponin I in muscle.
In vertebrate skeletal muscle, contraction is initiated by the elevation of the intracellular Ca2+ concentration. The binding of Ca2+ to TnC induces a series of conformational changes which ultimately release the inhibition of the actomyosin ATPase activity by Tnl. In this study we have characterized the dynamic behavior of TnC and Tnl in solution, as well as in reconstituted fibers, using EPR and ST-EPR spectroscopy. Cys98 of TnC and Cys133 of Tnl were specifically labeled with malemide spin label (MSL) and indane dione nitroxide spin label (InVSL). In solution, the labeled TnC and Tnl exhibited fast nanosecond motion. MSL-TnC is sensitive to cation binding to the high affinity sites (tau r increases from 1.5 to 3.7 ns), InVSL-TnC s sensitive to the replacement of Mg2+ by Ca2+ at these sites (tau r increase from 1.7 to 6 ns). Upon reconstitution into fibers, the nanosecond mobility is reduced by interactions with other proteins. TnC and Tnl both exhibited microsecond anisotropic motion in fibers similar to that of the actin monomers within the filament. The microsecond motion of TnC was found to be modulated by the binding of Ca2+ and by cross-bridge attachment, but this was not the case for the global mobility of Tnl.
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Department of Biological Sciences, Florida State University, Tallahassee 32306, USA.
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Fajer
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D000818:Animals; D003497:Cyclic N-Oxides; D004578:Electron Spin Resonance Spectroscopy; D007189:Indans; D009119:Muscle Contraction / Q000502:physiology; D018482:Muscle, Skeletal / Q000737:chemistry* / Q000378:metabolism; D009210:Myofibrils / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011817:Rabbits; D013113:Spin Labels; D019209:Troponin C / Q000737:chemistry / Q000378:metabolism*; D019210:Troponin I / Q000737:chemistry / Q000378:metabolism*
D003497:Cyclic N-Oxides; D007189:Indans; D013113:Spin Labels; D019209:Troponin C; D019210:Troponin I; C003028:N-(1-oxyl-2,2,6,6-tetramethyl-4-piperidinyl)iodoacetamide; C001445:1,3-indandione